CN102600461B - Give outer body ripe dendritic cells treatment tumour - Google Patents
Give outer body ripe dendritic cells treatment tumour Download PDFInfo
- Publication number
- CN102600461B CN102600461B CN201210027703.4A CN201210027703A CN102600461B CN 102600461 B CN102600461 B CN 102600461B CN 201210027703 A CN201210027703 A CN 201210027703A CN 102600461 B CN102600461 B CN 102600461B
- Authority
- CN
- China
- Prior art keywords
- cell
- dendritic cells
- composition
- bcg
- tumour
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 210000004443 dendritic cell Anatomy 0.000 title claims abstract description 171
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 127
- 238000011282 treatment Methods 0.000 title claims abstract description 18
- 210000004027 cell Anatomy 0.000 claims abstract description 95
- 230000036961 partial effect Effects 0.000 claims abstract description 56
- 238000000034 method Methods 0.000 claims abstract description 42
- 239000000427 antigen Substances 0.000 claims abstract description 38
- 108091007433 antigens Proteins 0.000 claims abstract description 38
- 102000036639 antigens Human genes 0.000 claims abstract description 38
- 230000004041 dendritic cell maturation Effects 0.000 claims abstract description 12
- 230000008569 process Effects 0.000 claims abstract description 11
- 210000001165 lymph node Anatomy 0.000 claims abstract description 10
- 102000013462 Interleukin-12 Human genes 0.000 claims abstract description 7
- 108010065805 Interleukin-12 Proteins 0.000 claims abstract description 7
- 230000006698 induction Effects 0.000 claims abstract description 5
- 229960000190 bacillus calmette–guérin vaccine Drugs 0.000 claims description 45
- 239000000203 mixture Substances 0.000 claims description 29
- 239000003104 tissue culture media Substances 0.000 claims description 22
- 239000003795 chemical substances by application Substances 0.000 claims description 20
- 238000012545 processing Methods 0.000 claims description 11
- 210000004369 blood Anatomy 0.000 claims description 8
- 239000008280 blood Substances 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims description 8
- 102000002689 Toll-like receptor Human genes 0.000 claims description 7
- 108020000411 Toll-like receptor Proteins 0.000 claims description 7
- 230000002779 inactivation Effects 0.000 claims description 7
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 6
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 claims description 6
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 claims description 6
- 238000002512 chemotherapy Methods 0.000 claims description 6
- 230000000139 costimulatory effect Effects 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 239000002158 endotoxin Substances 0.000 claims description 5
- 210000004700 fetal blood Anatomy 0.000 claims description 5
- 229920006008 lipopolysaccharide Polymers 0.000 claims description 5
- 238000001959 radiotherapy Methods 0.000 claims description 5
- 102000008070 Interferon-gamma Human genes 0.000 claims description 4
- 108010074328 Interferon-gamma Proteins 0.000 claims description 4
- 230000001939 inductive effect Effects 0.000 claims description 4
- 229960003130 interferon gamma Drugs 0.000 claims description 4
- 210000000952 spleen Anatomy 0.000 claims description 4
- 238000003786 synthesis reaction Methods 0.000 claims description 4
- RXNTZIJLOZMJTM-UHFFFAOYSA-N 2-(2h-quinolin-1-yl)ethanol Chemical compound C1=CC=C2N(CCO)CC=CC2=C1 RXNTZIJLOZMJTM-UHFFFAOYSA-N 0.000 claims description 3
- 101000599852 Homo sapiens Intercellular adhesion molecule 1 Proteins 0.000 claims description 3
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 claims description 3
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 3
- 239000002773 nucleotide Substances 0.000 claims description 3
- 125000003729 nucleotide group Chemical group 0.000 claims description 3
- 108091033319 polynucleotide Proteins 0.000 claims description 3
- 102000040430 polynucleotide Human genes 0.000 claims description 3
- 210000001541 thymus gland Anatomy 0.000 claims description 3
- 230000003827 upregulation Effects 0.000 claims description 3
- FQYRLEXKXQRZDH-UHFFFAOYSA-N 4-aminoquinoline Chemical compound C1=CC=C2C(N)=CC=NC2=C1 FQYRLEXKXQRZDH-UHFFFAOYSA-N 0.000 claims description 2
- -1 IL- 10 Proteins 0.000 claims description 2
- 108090001005 Interleukin-6 Proteins 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 210000004324 lymphatic system Anatomy 0.000 claims description 2
- 230000005809 anti-tumor immunity Effects 0.000 claims 1
- 230000017074 necrotic cell death Effects 0.000 claims 1
- 210000001744 T-lymphocyte Anatomy 0.000 abstract description 9
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 4
- 230000005975 antitumor immune response Effects 0.000 abstract description 2
- 230000028327 secretion Effects 0.000 abstract 1
- 239000002243 precursor Substances 0.000 description 47
- 241000699666 Mus <mouse, genus> Species 0.000 description 26
- 239000011159 matrix material Substances 0.000 description 23
- 206010036590 Premature baby Diseases 0.000 description 15
- 238000002347 injection Methods 0.000 description 15
- 239000007924 injection Substances 0.000 description 15
- 210000004881 tumor cell Anatomy 0.000 description 14
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 241001465754 Metazoa Species 0.000 description 12
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 10
- 210000001616 monocyte Anatomy 0.000 description 10
- 239000002953 phosphate buffered saline Substances 0.000 description 10
- 201000011510 cancer Diseases 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 230000035800 maturation Effects 0.000 description 9
- 238000000926 separation method Methods 0.000 description 9
- 102000004388 Interleukin-4 Human genes 0.000 description 8
- 108090000978 Interleukin-4 Proteins 0.000 description 8
- 229940028885 interleukin-4 Drugs 0.000 description 8
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 7
- 210000001015 abdomen Anatomy 0.000 description 7
- 230000028993 immune response Effects 0.000 description 7
- 230000003308 immunostimulating effect Effects 0.000 description 7
- 230000004614 tumor growth Effects 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 6
- 102000003816 Interleukin-13 Human genes 0.000 description 6
- 108090000176 Interleukin-13 Proteins 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 5
- 239000012980 RPMI-1640 medium Substances 0.000 description 5
- 230000004069 differentiation Effects 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 239000004033 plastic Substances 0.000 description 5
- 229920003023 plastic Polymers 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 102100035793 CD83 antigen Human genes 0.000 description 4
- 206010009944 Colon cancer Diseases 0.000 description 4
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 description 4
- 102100022297 Integrin alpha-X Human genes 0.000 description 4
- 102000013691 Interleukin-17 Human genes 0.000 description 4
- 108050003558 Interleukin-17 Proteins 0.000 description 4
- 239000012190 activator Substances 0.000 description 4
- 230000001464 adherent effect Effects 0.000 description 4
- 208000029742 colonic neoplasm Diseases 0.000 description 4
- 238000009295 crossflow filtration Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000011521 glass Substances 0.000 description 4
- 230000001506 immunosuppresive effect Effects 0.000 description 4
- 238000009169 immunotherapy Methods 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000002601 intratumoral effect Effects 0.000 description 4
- 210000002751 lymph Anatomy 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 108700028369 Alleles Proteins 0.000 description 3
- 102100036301 C-C chemokine receptor type 7 Human genes 0.000 description 3
- 101150013553 CD40 gene Proteins 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Natural products CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 101000716065 Homo sapiens C-C chemokine receptor type 7 Proteins 0.000 description 3
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 description 3
- BAQCROVBDNBEEB-UBYUBLNFSA-N Metrizamide Chemical compound CC(=O)N(C)C1=C(I)C(NC(C)=O)=C(I)C(C(=O)N[C@@H]2[C@H]([C@H](O)[C@@H](CO)OC2O)O)=C1I BAQCROVBDNBEEB-UBYUBLNFSA-N 0.000 description 3
- 102100039360 Toll-like receptor 4 Human genes 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 3
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- 230000002146 bilateral effect Effects 0.000 description 3
- 150000001768 cations Chemical class 0.000 description 3
- 239000002458 cell surface marker Substances 0.000 description 3
- 230000004087 circulation Effects 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 229960000554 metrizamide Drugs 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 210000005259 peripheral blood Anatomy 0.000 description 3
- 239000011886 peripheral blood Substances 0.000 description 3
- 239000000902 placebo Substances 0.000 description 3
- 229940068196 placebo Drugs 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 238000002271 resection Methods 0.000 description 3
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 108010050904 Interferons Proteins 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 101000746372 Mus musculus Granulocyte-macrophage colony-stimulating factor Proteins 0.000 description 2
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 2
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 230000005867 T cell response Effects 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N aldehydo-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- 230000000735 allogeneic effect Effects 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000010241 blood sampling Methods 0.000 description 2
- 210000002798 bone marrow cell Anatomy 0.000 description 2
- 238000002725 brachytherapy Methods 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000002648 combination therapy Methods 0.000 description 2
- 238000004891 communication Methods 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 238000000432 density-gradient centrifugation Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 102000054766 genetic haplotypes Human genes 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 229940031551 inactivated vaccine Drugs 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 238000007912 intraperitoneal administration Methods 0.000 description 2
- 210000002664 langerhans' cell Anatomy 0.000 description 2
- 210000003563 lymphoid tissue Anatomy 0.000 description 2
- 239000003607 modifier Substances 0.000 description 2
- 229940031346 monovalent vaccine Drugs 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 230000004936 stimulating effect Effects 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 239000007929 subcutaneous injection Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000013268 sustained release Methods 0.000 description 2
- 239000012730 sustained-release form Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- KMZHZAAOEWVPSE-UHFFFAOYSA-N 2,3-dihydroxypropyl acetate Chemical compound CC(=O)OCC(O)CO KMZHZAAOEWVPSE-UHFFFAOYSA-N 0.000 description 1
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000009410 Chemokine receptor Human genes 0.000 description 1
- 108050000299 Chemokine receptor Proteins 0.000 description 1
- 235000019743 Choline chloride Nutrition 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 102000016911 Deoxyribonucleases Human genes 0.000 description 1
- 108010053770 Deoxyribonucleases Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000360590 Erythrites Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 102000006354 HLA-DR Antigens Human genes 0.000 description 1
- 108010058597 HLA-DR Antigens Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Chemical compound OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000589343 Methylobacter luteus Species 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- UKEUCTXQMDFYAV-UHFFFAOYSA-N NC1=CC=NC2=CC=CC=C12.C1=CC=C2N=CC=CC2=C1 Chemical compound NC1=CC=NC2=CC=CC=C12.C1=CC=C2N=CC=CC2=C1 UKEUCTXQMDFYAV-UHFFFAOYSA-N 0.000 description 1
- 241000009328 Perro Species 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 206010064390 Tumour invasion Diseases 0.000 description 1
- 239000011358 absorbing material Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 230000009400 cancer invasion Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 description 1
- 229960003178 choline chloride Drugs 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical class OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 1
- 239000008119 colloidal silica Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000004940 costimulation Effects 0.000 description 1
- 230000002338 cryopreservative effect Effects 0.000 description 1
- 108010057085 cytokine receptors Proteins 0.000 description 1
- 102000003675 cytokine receptors Human genes 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 210000004544 dc2 Anatomy 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 235000003869 genetically modified organism Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000011173 large scale experimental method Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000001926 lymphatic effect Effects 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000873 masking effect Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000002966 oligonucleotide array Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000004091 panning Methods 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 210000003240 portal vein Anatomy 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 210000001147 pulmonary artery Anatomy 0.000 description 1
- 210000003492 pulmonary vein Anatomy 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002627 tracheal intubation Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000003954 umbilical cord Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/31—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterized by the route of administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
- A61K2239/50—Colon
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4615—Dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/462—Cellular immunotherapy characterized by the effect or the function of the cells
- A61K39/4622—Antigen presenting cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0639—Dendritic cells, e.g. Langherhans cells in the epidermis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
- C12N5/16—Animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/72—Undefined extracts from bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/05—Adjuvants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/24—Interferons [IFN]
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Cell Biology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Mycology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Oncology (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Hospice & Palliative Care (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The present invention provides the cell colony for including Partial mature dendritic cells that can be used for tumour individual administration.The dendritic cells of the Partial mature of about 1 about 10 hour or more long are acted on dendritic cell maturation reagent, tumour antigen, and full maturity effectively can be absorbed and process in tumor locus, at the lymph node that then can migrate to treatment individual.Once reaching in lymph node, full ripe antigen offers the appropriate cell factor (such as TNF α and IL 12) of dendritic cells secretion, and the follow-up anti-tumor immune response of induction is acted on T cell.
Description
The application is Application No. 200380108975.6, and the applying date is the division of the application of the same name on December 5th, 2003
Application.
Related application
This application claims the priority of U.S. Provisional Application filed in 6 days December in 2002 60/431,267, draw herein
Enter entire contents as reference.
Background of invention
Dendritic cells (DCs) are considered as the preferred carrier of tumour active immunotherapy.Animal experiment shows, based on DC's
Immunotherapy, which has, protects mice against the potentiality that generation tumour has formed tumour with elimination.This success reappears at least in part
In small-scale human clinical trial.Tested from small-scale security or theoretical validation, to illustrating activity or validity
The transition of large-scale experiment, is stagnated and (is seen below) due to the arduous and heavy of DC preparations.Therefore, although this product has
There is very big treatment potentiality, few companies are interested in developing the tumor vaccine based on DC.
Intra-tumor (IT) injection DCs is a kind of immunotherapy based on DC of special shape.After injection, DCs from apoptosis or
Antigen is absorbed in dying tumour cell, and moves to lymph node and offers antigen to T cell.Sent out actually in animal model
Validity (Cand ido et al., the Cancer Res.61 related to the apoptosis degree of tumour of existing this treatment：228-236,
2001), it shows this method with using chemotherapy or radiation therapy treatment tumour fully compatible before injection DCs.In addition,
There are several groups to prove, this therapeutic alliance to the tumour that has been formed especially effectively (Nikitina et al.,
Int.J.Cancer 94：825-833,2001；Tanaka et al., Int.J.Cancer 101：265-269,2002；Tong etc.
People, Cancer Res.61：7530-7535,2001).
Because tumour cell is the source of antigen, need to select and prepare tumour antigen before intra-tumoral injection, just as mesh
It is preceding they applied in most of external methods treated based on DC as.The selection of tumour antigen is usually by company to proprietary
The requirement of status is driven, and whether a small number of tumour antigens differentiated so far can provide significant clinical efficacy and need
Prove.In addition, univalent vaccine is generally produced using this tumour antigen, if tumour cell lowers the expression water of immunizing antigen
Flat, univalent vaccine may lose its effect.Certainly, tumour is manufactured under conditions of good manufacturing practive(GMP) (GMP) requirement is met
The requirement of antigen is traditional based on adding extra-pay on DC immunization methods.
DC s intra-tumoral injection makes dendritic cells be in immunosuppressive tumor environment.Known cancer produce cell because
Son, it inactivates DC s or t cell responses can be made to be inclined to the less Th2- types reaction of effect.Several research groups are repaiied using heredity
The DCs of decorations, to overcome these Inhibitory Effects, particular by the generation (IL-12 of cytokine interleukin element 12；Nishioka
Et al., Cancer Res.59：4035-4041,1999；Melero etc., Gene Therapy 6：1779-1784,1999) or table
Up to CD40 aglucons (Kikuchi et al., Blood 96：91-99,2000).The result of these groups description is encouraging, further
Intra-tumoral injection DCs is indicated as a kind of feasibility for the treatment of method.
Triozzi et al. (Cancer 89：2647-2654,2000) describe metastasis melanin tumor or patient with breast cancer
DCs IT injection.They obtain the result of tumor regression in 4 melanoma patients and 2 patients with mastocarcinoma.Disappear disease
The T cell of stove biopsy display infiltration, shows that DC actually have activated the immune response for tumour cell.It is all this
Injection DCs is feasible in a little as shown by data human tumors, can provide significant clinical efficacy.But, it has been observed that MHC
II classes antigen and B7-2 costimulatory moleculeses have obvious downward to the DCs of injection.The downward of these key molecules will be reduced
DCs immunostimulatory potential, but as provided by the present invention, this can be avoided by the Partial mature of DCs before administration.
DCs is present in peripheral tissues in prematurity form, prepares intake and process antigen.Exactly these prematurities are thin
Born of the same parents, in the presence of GM-CSF and IL-4, are most closely simulated by the DCs produced by monocyte.A variety of stimulations can start DCs's
Cell loses it and effectively absorbs the ability of antigen in maturation, maturation, and obtains its T cell stimulatory function.This process is
Complicated, at least to continue to complete for 48 hours in vitro.Another ripe result is that cell migration properties occur
Change.For example, some chemokine receptors of Maturation induction, including CCR7, CCR7 guide cell to the T cell area for flowing to lymph node
Domain, the ripe DCs activation T cell in the region, resistance I classes and II class MHC molecules are offered to the antigen on DC surfaces.
Inducing tolerance intersects (itself) antigen offered relevant (Kurts et al., J.Exp.Med.186 with DCs：239-
245,1997), hypothesis primarily now assumes that immature DC constantly offers antigen to immune system, to maintain tolerance
(Kurts et al., J.Exp.Med.184：923-930,1996), it is believed that cylinder mature DCs is needed to produce effective immune thorn
Swash.Clear and definite DC it is ripe there may be immunostimulation or inhibitive ability of immunity DCs (Chakraborty et al.,
Clin.Immunol.94：88-98,1999).Not yet illustrating now causes ripe stimulant any in two kinds of results
Definite property, but generally receive immunostimulation DCs generation IL-12 and express CD40 aglucons (Albert et al., Nature
Immunol.2：988-989,2001；The Supp l.EHA 4 of Lanzavecchia, Haematologica 84：23-25,
1999).Therefore, the DC of Partial mature should can be used for IT injections, particularly when the known thorn for making immature dendritic cell ripe
When sharp thing can cause the expression of CD40 aglucons and IL-12 generation.
Summary of the invention
The present invention provides a kind of method, for producing anti-tumor immune response, including give containing outer body into
The cell colony of ripe dendritic cells, so that individual is upon administration, the dendritic cells of Partial mature retain intake tumour antigen
Ability simultaneously can induce immune response.Although the overall immunosuppressive environment of intra-tumor, this method allows in tumour and tumour bedding
(bed) effective intake and processing in tumour antigen.It is this immune due to the negative effect to dendritic cells immunostimulating
The function of dendritic cells can be hindered by suppressing environment.Moreover, the dendritic cells of tumor maturation can not be given, because the cell can not have
Effect ground process antigen.
The dendritic cells of Partial mature can be administered directly to already present intra-tumor.Moreover, tumour bedding can be administered to, swollen
Tissue regions inside and around knurl, can treatment be surgical operation remove or tumor resection before or after carry out, chemotherapy,
Carried out prior to, concurrently with, or after radiotherapy or its combined therapy, etc..The dendritic cells of Partial mature can also be administered to direct stream
To tumour or the lymph node or lymphatic districts of peritumoral area.In addition, the dendritic cells of Partial mature can be administered to directly is
Tumour or tumour bedding or tumor afflicted organ or conveyed blood or the systemic vascular or lymph of lymph with tumour
In region.Further, the dendritic cells of Partial mature can generally be administered to circulation or lymphatic system, so that cell can be transported
To tumor region.
Available dendritic cells of the invention or its precursor, can be from such as skin, spleen, marrow, thymus gland, lymph node, umbilical cord
Obtained in blood or peripheral blood etc..In addition, the health that dendritic cells can match from individual to be treated or with individual HLA to be treated
Body is obtained.Before being contacted with maturing agent and being configured to the composition for individual administration, dendritic cells or its precursor can be with cold
Freeze and preserve.
Dendritic cells for the inventive method are Partial matures in vitro.For the appropriate of inducing dendritic cell maturation
Reagent includes, for example, but not limited to, BCG vaccine (BCG), interferon gamma (I FN γ), lipopolysaccharides (LPS), tumor necrosis factor α
(TNFα)；Imidazoquinolie compounds, such as imidazoquinolie -4- amines, such as 4- amino -2- ethoxyl methyls-α, α -
Dimethyl -1H- imidazos [4,5-c] -1- ethanol (being named as R848) or 1- (2- methyl-propyls) -1H- imidazos [4,5-c] quinoline
Quinoline -4- amine, and its derivative (WO00/47719 is fully incorporated by reference herein), the double-strand polyribonucleotide of synthesis, example
Such as poly- [I]：Poly- [C (12) U], etc.；Toll-like receptor (TLR) activator, such as TLR-3, TLR-4, TLR-7 and/or TLR-
9；Containing nucleotide sequence of non A-G hepatitis ripe known induction DC etc., or its combination.
BCG for the present invention may include complete BCG, BCG cell wall constituents, BCG- sources
Lipoarabidomannans, or any other components of BCG.In certain embodiments, BCG is heat inactivation BCG or formal
Woods handles BCG.Although alternatively BCG can be used alone, general BCG and IFN γ are used in combination and carry out inducing dendritic cell
Partial mature.The general every milliliter of tissue culture medium (TCM) of BCG effective dose is about 105-107Cfu, the effective dose of IFN γ is general every milliliter
Tissue culture medium (TCM) is about 100-1000 units.Dendritic cells activator is imidazoquinolie R898 in another embodiment.
TLR-4 activators R898 effective dose, usually using about 1-50 μ g/ml, is more often 5-10 μ with every milliliter of tissue culture medium (TCM)
g.Can be used alone imidazoquinolie, and also it can be used in combination with the BCG and/or IFN γ of such as effective dose.
Present invention additionally comprises composition, the compound is included, under dendritic cell maturation agent existence condition outer body into
Ripe dendritic cells, physiologically acceptable carrier, buffer solution and/or excipient in combination, for suffering from tumour individual administration.
Partial mature dendritic cells in the present invention used by composition generally raise costimulatory molecules such as, but not limited to CD80, CD86
Or CD54 expression.The cell can be expressed or not expression cell surface marker CD83, but remain to intake and process antigen.It is described
Composition generally includes about 102-1010, may also comprise the ripe dendritic cells of more parts.Before administration individual, dendritic cells portion
Be divided into it is ripe after can freezen protective for a period of time.Cell can be separated out of tumor patient body, be included for preparing the present composition
Partial mature BMDC, or separate cell from the patient HLA individuals matched, the dendron for preparing Partial mature
Cell.
Detailed description of the invention
Dendritic cells are a variety of antigen presenting cells found in a variety of lymphs and non-lymphoid tissue.(see Liu,
Cell, 106：259-62(2001)；Steinman, Ann.Rev.Immunol.9：271-96(1991)).Dendritic cells include spleen
Masking (veiled) cell in lymphoid dendritic cell, epidermis Langerhans' cells and blood circulation.In general, according to it
Form, surface II classes MHC high level expression and lack some other T cells, B cell, monocyte and NK
The surface marker of expression divides dendron cell mass.Specifically, (also referred to as monokaryon is thin for the dendritic cells in monocyte-source
Born of the same parents' dendritic cells) CD11c, CD80, CD86 are often expressed as, it is HLA-DR+, but be CD14-。
On the contrary, the precursor (generally monocyte) of monocytic dendritic cells is usually CD14+.Monocytic dendritic cell is thin
The precursor of born of the same parents can have their tissue from any, specifically lymphoid tissue, such as obtained in spleen, marrow, lymph node and thymus gland
.The precursor of monocytic dendritic cells can also be separated from the circulatory system.Peripheral blood is also monocytic dendritic cell precursor
The source that is easy to get.Cord blood is another source of monocytic dendritic cell precursor.Monokaryon can be separated from a variety of organisms
Immune response in the precursor of cells Dendritic cells, wherein organism can trigger.This kind of organism includes animal, for example, wrap
Include the mankind and non-human animal, such as primate, mammal (including dog, cat, mouse and rat), birds (including chicken)
And its genetically modified organism.
In certain embodiments, from health volunteer or it can need in subject's body of immunostimulation, separation monokaryon is thin
The precursor of born of the same parents' dendritic cells and/or immature dendritic cells, such as that tumor patient or may benefit from or need cell to exempt from
Other subjects (i.e. the subject of bacterium or virus infection etc.) that epidemic disease is stimulated.Dendritic cell precursor and/or immature tree
Prominent cell can also be obtained from the HLA- healthy individuals matched, for partial activation and be administered to and needed the HLA- of immunostimulation to match
Subject.
Dendritic cell precursor and immature dendritic cells
It is known in the art from a variety of sources include blood and marrow in separation rich in dendritic cell precursor and prematurity dendron
The method of the cell colony of cell.For example, can be removed by collecting anticoagulant heparin blood, single blood sampling composition art or leucocyte
Method, buffy coat is prepared, rosette, centrifugation, density gradient centrifugation is formed and (for example uses(for example
FICOLL-)、(colloidal silica particles (diameter 15-30nm) are coated with non-parsing polyvinylpyrrolidone
(PVP)), sucrose etc.), differential lysis of cells, filtering etc., separation dendritic cell precursor and prematurity dendritic cells.In some realities
Apply in scheme, can defibrinate former to remove blood platelet and splitting erythrocyte, prepare white for example, by collecting subject's blood
Cell mass.Can be for example, byThe methods such as gradient centrifugation, antibody elutriation (panning), optionally for monokaryon
Dendritic cell precursors and be enriched with dendritic cell precursor and prematurity dendritic cells.
Dendritic cell precursor and prematurity dendritic cells can be optionally prepared in closed sterile system.It is used herein
Term " closed sterile system " or " closed system " refer to reduce or eliminate its exposed to ambient non-sterile or circulation air or
Other non-sterile situations.Closed system for separating dendritic cell precursor and prematurity dendritic cells, is typically not included in top
Density gradient centrifugation in portion's opening test tube, outdoor cell transfer, in tissue culture plate or unsealing shaking flask cell training
Support etc..In a representative embodiment, closed system is allowed in be not exposed to non-sterile air in the case of, by dendron
Cell precursors and prematurity dendritic cells are transferred aseptically to from initial collection device in sealable tissue culture dishes.
Another is reported is for separating the method for dendritic cell precursor, using commercialization processing plastic matrix (for example
Globule or magnetic bead), the monocyte and other " non-dendritic cell precursors " of selective removal attachment are (see such as United States Patent (USP)
Numbers 5,994,126 and 5,851,756).The monocyte and non-dendritic cell precursor of attachment are discarded, retains non-adherent cell, is used
In in vitro culture and maturation.In another method, the cell culture of single blood sampling composition art adds thereto in plastic culture bag
Enter plastics i.e. polystyrene or styrene microcarrier bead to improve the surface area of sack.Cell fully cultivates a period of time, extremely
Some cell adherences wash away non-adherent cell on pearl from bag.(Maffei et al., Transfusion 40：1419-
1420(2000)；WO 02/44338, is hereby incorporated by reference).
In some other embodiments, monocytic dendritic cell precursor by with monocyte binding matrix adhesion and
Separation, as described in WO 03/010292, the disclosure of which is incorporated herein by reference.For example, leucocyte group is (such as by white thin
Born of the same parents remove method separation) it can be contacted with the adhering substrate of monocytic dendritic cell precursor.When leucocyte group contacts with matrix,
Monocytic dendritic cell precursor in leukocyte population is preferentially adhered in matrix.Other leucocytes (including other potentialities
Dendritic cell precursor) reduction of binding matrix affinity, so as to allow the dendritic cell precursor priority enrichment of monocyte in matrix
On surface.
Appropriate matrix includes, those larger matrix of the ratio between such as surface area and volume.Matrix can be for example micro-
Grain or fibrous matrix.Appropriate matrix of microparticles includes, for example the coated plastic particles of glass granules, plastic particles, glass, glass
The coated ps particle of glass and other be suitable to adsorbed proteins bead.The fibrous matrix used suitable for the present invention includes
Microcapillary and microvillose membrane etc..Particulate or fibrous matrix are not generally reducing the situation of adherent cell viability substantially
Under, allow to elute the monocytic dendritic cell precursor of adhesion.Particulate or fibrous matrix can be it is substantially non-porous,
To elute monocytic dendritic cell precursor or dendritic cells from matrix." substantially non-porous " matrix refer to present at least at
Most of stomata in matrix is smaller than cell, so that the cell of Medium Culture trapping minimize.
By adding binding medium, optionally strengthen the adhesion of monocytic dendritic cell precursor and matrix.Appropriate
Binding medium includes monocytic dendritic cell precursor culture medium (such as AIM-RPMI 1640, DMEM, X-VIVO
Etc.), supplement individually or in any combination such as cell factor (such as granulocyte/macrophage colony stimulatory factor (GM-
CSF), interleukin-4 (IL-4), or interleukin-13 (IL-13))；Blood plasma, serum (such as human serum, such as it is autologous or
Allogeneic serum)；The protein of purifying, such as seralbumin；Bivalent cation (such as calcium and/or magnesium ion)；And other have
Help the molecule that monocytic dendritic cell precursor and substrate specificity are adhered to, or prevent non-monocytic cells dendritic cell precursor with
The molecule of matrix attachment.In certain embodiments, blood plasma or serum heated can be inactivated.Heat inactivated blood plasma both can be
Can also be allosome with leucocyte consubstantiality.
After monocytic dendritic cell precursor is adhered in matrix, before non-adhering leucocyte and monocytic dendritic cells
Body/base complex is separated.Any appropriate method is used equally for the separation of non-adherent cell and compound.For example, can sink
The mixture of shallow lake non-adhering leucocyte and compound, removes or drains non-adhering leucocyte and medium supernatant.Alternatively,
The supernatant for including non-adhering leucocyte can be removed or drained with centrifugal mixture, it is separated with sediment composite.
In another method, can from the cell mass rich in leucocyte isolating monocytic dendritic cell precursors, wherein in vain
Cell prepared using tangential flow filtration device (on June 19th, 1 applies, described in international patent PCT/US03/19428,
It is herein incorporated by reference).Tangential flow filtration device for separating the cell colony rich in monocytic dendritic cell precursor,
It can include and remove unit, filtrate chamber and filter positioned there between with interaction flowing (cross-flow) room.Filter exists
Side retains surface and interaction flow chamber carries out fluid communication, filters surface in opposite side and filtrate chamber carries out fluid communication.Hand over
Mutual flow chamber has entrance, suitable for the blood constituent Sample introduction including leucocyte is interacted into flow chamber, is put down with filter retention face
OK.Meanwhile, interaction flow chamber provides outlet, is placed between two parties in groove and faces the part that filter retains surface.It is suitable for tangential
Flow normally about 1-10 microns of the filter average pore size of filter.Filter average pore size can be about 3-7 microns.It may also include to set
Random sample product ingress rate provides the device for entering cross flow one chamber inlet, and control filtrate enters the mistake of filtrate chamber by filter
The device of filtering velocity degree.The rate of filtration control device limitation rate of filtration is less than the non-confrontation rate of filtration of filter.Including blood group
Sample including point, can be by source device such as leukapheresis device, or contains the sample collected from leukapheresis device
The container of product is provided.
Monocytic dendritic cell precursor and for precursor be enriched with cell mass, can cultured in vitro to break up, Partial mature
And/or amplification.Prematurity dendritic cells, dendritic cell precursor and the other cells of separation used herein, refer to manually locate
The cell that reason is separately present with its natural surroundings, and unnatural products.The cell of separation can be purified, semipurified form is deposited
, or under non-natural environment.In short, ex vivo differentiation is commonly included in the presence of one or more differentiation agents,
Culture of monocytic dendritic cell precursors or the cell mass with dendritic cell precursor.Appropriate differentiation agent may include, for example carefully
The intracellular growth factor (such as cell factor such as (GM-CSF), interleukin-4 (IL-4), interleukin-17 (IL-7), leucocyte
Interleukin 13 (IL-13) and/or its combination).In certain embodiments, to be differentiated to form monokaryon thin for monocytic dendritic cell precursor
The prematurity dendritic cells in born of the same parents source.
Dendritic cell precursor can be cultivated and broken up under appropriate condition of culture.Appropriate tissue culture medium (TCM) includes,
But it is not limited to, AIM-RPMI1640, DMEM, X-Etc..Serum, amino can be supplemented into tissue culture medium (TCM)
Acid, vitamin, cell factor such as GM-CSF and/or IL-4, IL-7, IL-13, bivalent cation etc., to promote point of cell
Change.In certain embodiments, dendritic cell precursor can be incubated in serum free medium.Condition of culture can optionally exclude any
The product of animal origin.Be typically used for dendritic cell culture media combination of cytokines include GM-CSF and IL-4, IL-7 or
IL-13, every kind of about 500 units/ml.When dendritic cell precursor is differentiated to form immature dendritic cells, phenotype is similar to skin
Langerhans' cells.Immature dendritic cells are generally CD14-And CD11c+, low expression level CD86 and CD83, and can pass through
Special endocytosis capture soluble antigen.
Dendritic cell maturation agent may include, be such as, but not limited to, BCG, IFN γ, LPS, TNF α, imidazoquinolie chemical combination
Thing, such as imidazoquinolie -4- amines such as 4- amino -2- ethoxyl methyl-α, alpha-alpha-dimethyl -1H- imidazos [4,5-c]
Quinoline -1- ethanol (be referred to as R848) or 1- (2- methyl-propyls) -1H- imidazos [4,5-c] quinolin-4-amines, and they spread out
Biological (WO00/47719, herein incorporated by reference)；The double-strand polyribonucleotide of synthesis, such as poly- [I]：Poly- [C (12) U]
Deng；Toll-like receptor (TLR) activator, such as TLR-3, TLR-4, TLR-7 and/or TLR-9；Ripe comprising known induction DC
Nucleotide sequence of non-methylated CpG motif etc., or its any combination.It is big that BCG effective doses are generally every milliliter of tissue culture medium (TCM)
About 105-107cfu.IFN γ effective dose is generally every milliliter of tissue culture medium (TCM) about 100-1000U.BCG vaccine (BCG) is
M.bovis avirulent strain.Refer to complete BCG and cell-wall components, BCG sources for BCG herein
Lipoarabidomannans and other BCG components.BCG is optionally at inactivation, such as heat inactivation BCG, formalin
Manage BCG etc..Imidazoquinolie compounds such as imidazoquinolie -4- amines, such as 4- amino -2- ethoxyl methyls-α, α -
The use effective dose of dimethyl -1H- imidazos [4,5-c] quinoline -1- ethanol (being referred to as R848) is every milliliter of culture medium about 1-
50 μ g, more typically, every milliliter of culture medium use about 5-10 μ g.Imidazoquinolie compounds can be used alone, or with for example
BCG and/or IFN γ or other TLR agonist combinations are used.
General immature DC s and effective dose dendritic cell maturation agent, such as BCG and IFN γ effect about 1-10 are small
When.Full maturity typically at least needs to incubate 24 hours, more typically incubates about 48-72 hours.Immature dendritic cells
It can be cultivated under appropriate condition of culture and Partial mature.Appropriate tissue culture medium (TCM) includes：AIM-RPMI
1640、DMEM、X-Etc..Amino acid, vitamin, cell factor such as GM-CSF can be supplemented into tissue culture medium (TCM)
And/or IL-15, bivalent cation etc., to promote the maturation of cell.The GM- that typical combination of cytokines is 500 units/ml
CSF and 100ng/ml IL-15.
The Partial mature of prematurity dendritic cells can be monitored for method known to dendritic cells using this area.It can adopt
With it is well known that technology, such as flow cytometry, immunohistochemistry, to detect cell surface marker thing.It can also pass through
The generation (such as by ELI SA, other immunoassays or utilizing oligonucleotide arrays method) of cell factor is thin to monitor
Born of the same parents.In dendritic cell maturation agent, such as in the case of GM-CSF and IL-4 are present, according to present invention culture and the DCs of Partial mature
In, it can use it is well known that method determines the level of phosphorylation JAK2 (janus activates kinases 2), to indicate the initial of maturation.
The induced expression of cell surface marker thing and cell factor, and signaling molecule phosphorylation, such as jak2 are also regarded as tree
Prominent cell is suitable for administration to individual to induce the indicator of immune response.
Immature dendritic cells only ripe a period of time, so that dendritic cells Partial mature.Full ripe DCs loses
The ability of intake antigen and the expression for raising costimulation cell surface molecule and cytokine profiles.Specifically, into
The expression of ripe DCs MHCI classes and II class antigens is higher than immature dendritic cells, and ripe dendritic cells are generally identified as
CD80+、CD83+、CD86+And CD14-.MHC height expression cause DC surface antigens density increase, and costimulatory molecules CD80 and
CD86 up-regulation, by the homologue (counterpart) of costimulatory molecules, such as the CD28 in T cell makes T cell activation
Signal is enhanced.For the present invention Partial mature dendritic cells ,-as include once be exposed to dendritic cell maturation agent, table
Reveal costimulatory molecules --- include but is not limited to the dendritic cells of CD80, CD86 and/or CD54 up-regulated expression.The cell can
Whether expression CD83 or not, but keeps the ability of intake and process antigen.In addition, the dendritic cells of Partial mature can produce TNF-
α, IL-6, IL-10 and/or IL-12, general prematurity dendritic cells will not produce these molecules.
Full ripe dendritic cells are not the preferred of the present invention, because they can not effective process antigen.In addition, being used for
Prematurity dendritic cells in existing method are nor required, because intra-tumor general nature formation immunosuppressive environment, bag
Include the cell factor for having notified the larger concentration for preventing prematurity dendritic cells antigen from processing.In the present invention, Partial mature
Prematurity dendritic cells lower the cytokine receptor on cell surface, reduce them to being present in the cell in inside tumor space
The sensitiveness or reactivity of the immunosuppressive effects of the factor, make cell effectively to process the tumour antigen for being present in intra-tumor.One
The dendritic cells of denier Partial mature are ripe in intra-tumor, its generation for example, by TNF-α and IL-12, chemokine receptors
CCR7 expression is determined, and cell can migrate at lymph node, and cell will be contacted with T cell there, and up-regulation is anti-to tumour
Former immune response.
According to another aspect of the present invention, given before maturation or after Partial mature before patient, DCs can be preserved,
Such as Cord blood.The cryopreservative that can be used, includes but is not limited to, dimethyl sulfoxide (DMSO), glycerine, polyvinyl pyrrole
Alkanone, polyethylene glycol, albumin, glucan, sucrose, ethylene glycol (ethylene glycol), i- erythrites, D-ribose alcohol, D-
Mannitol, D- D-sorbites, i- inositols, D- lactose, Choline Chloride, amino acid, methanol, acetamide, acetin and nothing
Machine salt.Cooling is very crucial at a slow speed for control.Different antifreezing agent and different cell types typically using it is different most
Good cooldown rate.The heat in molten bonding (fusion) stage that water build-ups ice should typically minimize.Cooling procedure is available, for example
Can programme controlled refrigerating plant or methanol bath procedure complete.Sequencing refrigerating plant allows to determine optimal cooldown rate, just
In standardization and recursive cooling.Programmable controlled-rate freezer unit, such as Cryomed or Planar, will can be freezed
Mode is adjusted to required cooling-rate curves.
After thorough freezing, the DCs of monocytic precursor cells, immature DCs or Partial mature can be transferred to length rapidly
In the Cord blood container of phase.In a typical embodiment, sample can Cord blood in liquid nitrogen (- 196 DEG C) or its steaming
In gas (- 165 DEG C).For candidate stem cell, specifically from marrow or the candidate stem cell of peripheral blood, the principle, operation,
Freezen protective and long-term preservation, are very suitable for the cell of the present invention.It, which is discussed, can be found in, such as following bibliography,
This is incorporated herein by reference：Taylor et al., Cryobiology27：269-78(1990)；Gorin, Clinics in
Haematology 15：19-48(1986)；Bone-Marrow Conservation, Culture and
Transplantation, Proceedings of a Panel, Moscow, Jul.22-26,1968, International
Atomic Energy Agency, Vienna, pp.107-186.
Preferably, frozen cell thaws (be for example held in 37-41 DEG C of water-bath) rapidly, is cooled down immediately after defrosting.Can
Can need processing cell in case thaw after cell agglutination.It a variety of methods can be used to prevent aggegation, include but is not limited to, before freezing
And/or DNase (the Cancer such as Spitzer 45 are added afterwards：3075-85 (1980)), low molecular weight dextran and citric acid
Salt, HES (Stiff etc., Cryobiology 20：17-24 (1983)), etc..If antifreezing agent is poisonous to human body
Property, it will should be removed before treatment use in its Partial mature DCs from defrosting.A kind of method for removing antifreezing agent is by it
It is diluted to insignificant concentration.The Partial mature DCs of freezing once thaw and recover, you can according to the part herein in regard to non-frozen into
Ripe DCs description is administered.
The dendritic cells of Partial mature are given in vivo
The invention provides the side for giving tumor patient Partial mature dendritic cells or cell colony rich in this cell
Method.In certain embodiments, by obtaining dendritic cell precursor or immature dendritic cells, in dendritic cell maturation agent,
These are thin for such as BCG and IFN γ, or any other as in the presence of maturing agent listed above, differentiation and Partial mature
Born of the same parents, to realize the above method.The dendritic cells of Partial mature can be used method and composition well known to those skilled in the art with
The acceptable carrier of physiology, excipient, buffer solution or diluent are prepared together.The dendritic cells of Partial mature can directly to
Need the patient of immunostimulation.General about 102-1010Cell be suspended in pharmaceutically acceptable carrier.Cell direct injection
Enter tumour, or be injected near tumour or tumour bedding, neighbouring or the circulation being in contact with tumour or tumour bedding or lymph area
Domain, to ensure that cell can be close to tumour antigen.
Such as, but not limited to, cell can be administered directly to intra-tumor, the tumour after surgical operation removing or tumor resection
Drainage (draining) lymph node, flow direction or supply tumour or the tumor invasion device directly contacted around bedding, tumour, with tumour
In the blood vessel or lymphatic vessel of official, such as portal vein or pulmonary vein or artery etc..For example chemotherapy or it can put in other oncotherapies
While treatment or afterwards, the dendritic cells of the Partial mature of the present invention are given.In addition, the dendritic cells of Partial mature of the present invention can
With other reagent co-administereds, the reagent plays promoter effect, makes maturing dendritic cell, and/or close or adjacent to tumour
Antigen in nearly tumor region is processed.In addition, dendritic cells can also be prepared or are mixed into sustained-release matrix, implantation tumour
Or region inside or around tumour bedding, so that cell is released slowly into inside tumour or tumour bedding, contacted with tumour antigen.
The dendritic cells of Partial mature of the present invention can be given using any formulation and mode for being suitable to administration.For example, cell
It can be combined with pharmaceutically acceptable carrier, can be using syringe, conduit, intubation etc. administration.As described above, cell can be formulated in
In sustained-release matrix.When being administered by this way, preparation can be administered by the method for the matrix being adapted in use to.It is other to be applied to this
The medication and mode of invention are well known to those skilled in the art.
The composition of the present invention itself can be used for individual treatment.In addition, also composition can be combined with any other method
Using treating tumour.For example, the inventive method can be with cancer surgery resection, chemotherapy (cytotoxic drug, apoptosis examination
Agent, antibody etc.), radiotherapy, cold therapy, brachytherapy, immunotherapy (give the Activates Dendritic of antigen specific mature
Cell, NK cells, specific for tumour antigen antibody etc.) etc. be applied in combination.Any of these methods all can also
It is used in any combination.Combined therapy can be carried out simultaneously or sequentially, any order of administration determined by treating physician.
In another embodiment, dendritic cells have identical MHC (HLA) haplotype with recipient subjects.It is determined that
The method of subject's HLA haplotypes is known in the art.In the relevant embodiments, the dendritic cells of Partial mature and acceptor by
Examination person is allosome.Variant cell typically matches with least one MHC allele (for example has at least one but not institute
There is MHC allele).In a less typical embodiment, dendritic cells and recipient subjects are entirely allosome each other
, but all at least one common MHC allele.
Embodiment
The following example is merely to illustrate various aspects of the invention, and should not be construed as in any way to the present invention's
Limit.
Embodiment 1
In this example, the monocytic dendritic cell precursor separated by tangential flow filtration, with being supplemented with 2% human serum
The X- of albumin (HSA) and 500U/ml GM-CSFUsing concentration as every milliliter 10 in blake bottle or cell bags6
Monocyte culture.After 5 days, the DC s from blake bottle and cell bags either leave and do not handle (iDCs) or in BCG
(1：400 dilutions) and interferon gamma (IFN γ, 500U/ml) (pmDC s) existence condition under Partial mature 4 hours.Collect cell,
Wash and mixed 48 hours with the PKH67-A549 tumour cells that equivalent radiofluorescence is marked.After incubation, cell is washed, algae red is used
The CD11c monoclonal antibody specifics of albumen (PE) mark are dyed, and through flow cytometry.It is provided in the number of table 1
Value represents the CD11c in DC+Cell, namely mark PKH67 antigen-positive cells (MFI=average fluorescent strengths) percentage
Than.Control includes the DCs before analysis just with PKH67-A549 mixing with cells.
Table 1.
These as shown by data, as measured by absorbing the cell percentages of antigen or the numerical value of tumour cell absorbing material,
In this experiment, the DCs of Partial mature is compared with immature dendritic cells, in terms of antigen uptake more preferably.
In another example, surgically remove after growth tumour, leukopheresis has been carried out to patient.Pass through
Tangential flow filtration purifying monocytes from leukopheresis product.The monocyte of purifying is broken up and led to as described above
Cross the BCG (5x10 fixed exposed to heat inactivation and formalin6Cfu/ml) IFN γ (500U/ml) Partial mature.It is formulated for
The dendritic cells of the Partial mature of administration, and individual tumors bedding is injected under ultrasound guidance.External test is found, from individual
The dendritic cells of separation, can the different in nature cytotoxic T cell response of induced tumor.Moreover, it has been found that the tumor recurrence time is blocked
Or be largely postponed.
Embodiment 2
In the present embodiment, human colon cancer cell is transplanted to mouse using well-known process, is allowed to produce heterograft and swells
Knurl.After tumour is formed, by animal packet, placebo, immature dendritic cells or part are given into every group of animal tumor
Ripe dendritic cells.Then determine the size of each group tumour and be compared.
In short, by 100,000 tumour cell of rib subcutaneous injection to the right, being generated with female Balb/c mouse
People's CT26 colon cancer tumours.Treatment starts from the 15th day, and now all mouse substantially carry tumour, and tumor size is
10mm2-45mm2。
According to standard scheme dendritic cells are prepared from female Balb/c mouse femurs marrow.In short, after erythrocyte splitting
Bone marrow cell and mouse GM-CSF (200U/ml) and I L-4 (200U/ml), in the RPMI 1640 of 10% hyclone is supplemented with
It is incubated 13-14 days, the fresh culture of factor-containing was added after the 3rd and the 7th day.Dendron for acquisition Partial mature is thin
Born of the same parents, expose cells to the inactivated vaccine (BCG) of 400 times of dilution and the mouse interferon (IFN γ) of every milliliter of 500 units
In, wherein BCG activity is every milliliter of 0.5-1x10 before inactivation6CFU.Cell incubation 8 hours, then with 14.5%
Metrizamide (metrizamide) layering and in 4 DEG C, 1750rpm is centrifuged 15 minutes.Interface cell is collected, is washed,
The frozen ground that can be survived in 10% dimethyl sulfoxide (DMSO) is stored in liquid nitrogen.After being thawed at 37 DEG C, cell is washed twice, resuspended, dense
Spend for every 50 microlitres of 1,000,000 cells.
Endoxan processing tumor-bearing mice (50mg/kg) is given in the 15th and 17 day intraperitoneal.The 16th, 17,18 and 21
BMDC is expelled to intra-tumor by it, and (the μ l of simulated injection 50 phosphate buffered saline (PBS), immature DC or maturation 8 are small
When DC).The next day determine tumour growth, survey 60 days.
In phosphate buffered saline (PBS) intra-tumor treatment group, the 40th day or before all 4 mouse tumors be all higher than
100mm2.On the contrary, the mice tumors grew that intra-tumor is handled with immature DC slows down, there are 2 tumours small in the 40th day 4 mouse
In 100mm2.There are 3 tumours to be less than 100mm in 4 mouse that 40th day is handled with Partial mature DC2, and have in these animals
2 at the 50th day still without obvious tumour.
Embodiment 3
In the present embodiment, human colon cancer cell is administered to by mouse using well-known process, is allowed to produce heterograft and swells
Knurl.After first time injection tumour cell, then give the tumour cell of one dosage of animal.After 15 days, by animal packet, to every
The dendritic cells of placebo, immature dendritic cells or Partial mature are given in group animal tumor.Each group is then determined to swell
The size of knurl is simultaneously compared.
In short, by 100,000 tumour cell of rib subcutaneous injection to the right, being generated with female Balb/c mouse
People's CT26 colon cancer tumours.Then at the 3rd day, rib injected 100,000 tumour cells to the left.Treatment starts from the 15th day, now
All right rib abdomens of mouse substantially carry tumour, and tumor size is 10mm2-45mm2。
According to standard scheme dendritic cells are prepared from female Balb/c mouse femurs marrow.In short, after erythrocyte splitting
Bone marrow cell and mouse GM-CSF (200U/ml) and I L-4 (200U/ml), in the RPMI 1640 of 10% hyclone is supplemented with
It is incubated 13-14 days, the fresh culture of factor-containing was added after the 3rd and the 7th day.Dendron for acquisition Partial mature is thin
Born of the same parents, expose cells to the mouse interferon (IFN γ) of 400 times of inactivated vaccines (BCG) diluted and every milliliter of 500 units
In, wherein BCG activity is every milliliter of 0.5-1x10 before inactivation6Colony forming unit.Cell then with 14.5% metrizamide
It is layered and in 4 DEG C, 1750rpm per minute is centrifuged 15 minutes.Interface cell is collected, is washed, and in 10% dimethyl sulfoxide
(DMSO) frozen ground that can be survived in is stored in liquid nitrogen.After being thawed at 37 DEG C, cell is washed twice, resuspended, and concentration is every 50 microlitre 1
Million cells.
Endoxan processing tumor-bearing mice (50mg/kg) is given in the 15th and 17 day intraperitoneal.The 16th, 17,18 and 21
My god, by dendritic cells be expelled to right rib abdomen intra-tumor (or the μ l of simulated injection 50 phosphate buffered saline (PBS), immature DC,
Ripe 8 hours DC or ripe 24 hours DC).The next day determine tumour growth, survey 60 days.
In phosphate buffered saline (PBS) intra-tumor treatment group, all 5 mouse were more than in the front right rib abdomen tumour of the 35th day
100mm2, 3/5 animal at the 40th day or front left rib abdomen tumour be more than 100mm2.On the contrary, treating intra-tumor with immature DC
The mice tumors grew on (only right side) slows down：One mouse is in the 60th day bilateral without 4 in obvious tumour, and 5 mouse
Only 100mm is respectively less than in the 40th day bilateral tumour2.There are 3 to the 50th day in 5 mouse with Partial mature DC processing in 8 hours
When tumour clear up completely.Further declined with tumour growth in the group of Partial mature DC processing in 24 hours.3 in 5 mouse
Cleared up completely in the 50th day bilateral tumour, the left rib abdomen tumour of an animal is cleared up completely, right rib abdomen tumor section is controlled (the 40th day
＜ 100mm2), animal both sides tumor section control (the 40th day both sides all ＜ 50mm2)。
Embodiment 4
In the present embodiment, allogeneic tumor is produced with mouse tumor with human tumor cells as described above.By by carefully
Born of the same parents combine the dendritic cell maturation agent of BCG and IFN γ exposed to imidazoquinolie R898 or R898, make the immature tree of differentiation
Prominent cellular portions are ripe.The dendritic cells, immature dendritic cells and placebo of Partial mature are given to intra-tumor, it is relatively more each
The size of group tumour.
In short, forming tumour with mouse according to embodiment 2.Dendritic cells are prepared according to embodiment 2 and 3, but
Partial mature cell is by exposing cells to 5 μ g/ml immune response modifier (dendritic cell maturation agent) R848, or 5 μ
In the BCG and interferon gamma of g/ml immune response modifier R848 and concentration described in embodiment 2 and 38 hours and obtain.
Processing method be the same as Example 3.The tumour growth of the mouse of phosphate buffered saline (PBS) intra-tumoral injection is not controlled.
Individually with 5 mouse of R848 Partial matures DC processing, there is a tumour growth to clear up completely, and combine and use R848 and BCG
In 5 mouse of IFN γ Partial mature DC intra-tumors processing, there are 3 tumour growths to clear up completely.In embodiment 3 and 4
9 mouse that tumour is cleared up completely, are attacked for about 30 days after tumor regression with 1,000,000 tumour cells in right rib abdomen.Do not move
There is detectable tumour growth in thing, shows the immanoprotection action to CT26 tumour cells.
Embodiment 5
In the present embodiment, the expression that the dendritic cells prepared in above example carry out surface molecular is examined and determine.In short,
Dendritic cells in mouse prepared by 2-4 is tested, by flow cytometry after fluorescent antibody staining, to examine and determine cell surface molecule
Expression.As a result represented with the average fluorescent strength of the percentage of surface marker Phenotype positive cell and certain surface marker, such as table 2
With 3.
Table 2
Table 3
Embodiment 7
After patient of the diagnosis with solid tumor is using chemotherapy, radiotherapy, cold therapy or brachytherapy treatment, intra-tumor
Give the dendritic cells of Partial mature.After treatment, tumor regression and it can prevent that patient tumors from recurring.
Above-described embodiment only for explanation, is not intended to limit the present invention and requires the scope of claim.To those skilled in the art
Other changes of the present invention are it is clear that and be contained in accessory claim.All publication cited herein, patent, specially
Profit application and other bibliography, its content are also incorporated by reference.
Claims (14)
1. a kind of composition, it is contained in the dendritic cells of the external Partial mature in the presence of dendritic cell maturation agent, and medicine
The acceptable carrier for vivo medicine-feeding on, wherein the dendritic cells of described Partial mature keep intake and process antigen
Ability, and compared with ripe dendritic cells, wherein the dendritic cells of described Partial mature produce TNF α, IL-6, IL-
10, and/or IL-12, and show costimulatory molecules CD80, CD86 and/or CD54 up-regulation,
Dendritic cell maturation agent therein is that BCG vaccine (BCG), BCG and interferon gamma (IFN γ), lipopolysaccharides (LPS), tumour are bad
Necrosis factor α (TNF α), imidazoquinolie compounds, the double-strand polybribonucleotide of synthesis, Toll-like receptor (TLR) excitement
Agent, the nucleotide sequence containing non-methylated CpG motif ripe known induction DC, or its any combination.
2. the composition of claim 1, dendritic cells therein are obtained from skin, spleen, marrow, thymus gland, lymph node, Cord blood or outer
All blood.
3. the composition of claim 2, wherein described dendritic cells are obtained from individual to be treated.
4. the composition of claim 2, dendritic cells therein are obtained from the healthy individuals matched with individual HLA to be treated.
5. the composition of claim 1, wherein the LAM that the BCG originates comprising complete BCG or BCG.
6. the composition of claim 1, wherein the BCG is the BCG of the heat inactivation or BCG of formalin processing.
7. the BCG of the composition of claim 1, wherein effective dose is every milliliter of tissue culture medium (TCM) 105-107Cfu, effective dose
IFN γ is every milliliter of tissue culture medium (TCM) 100-1000 unit.
8. the composition of claim 1, wherein described imidazoquinolie compounds are imidazoquinolie -4- amines.
9. according to claim 8 composition, wherein imidazoquinolie -4- amines be 4- amino -2- ethoxyl methyls-α, α -
Dimethyl-lH- imidazos [4,5-c] quinoline -1- ethanol, or 1- (2- methyl-propyls) -1H- imidazos [4,5-c] quinoline -4-
Amine, or derivatives thereof.
10. the composition of claim 1, the double-strand polyribonucleotide of synthesis therein is poly- [I]：Poly- [C (12) U].
11. any one of claim 1-10 composition, which part mature dendritic cell is used as radiotherapy, chemotherapy or its combination
Assistant agent apply.
12. any one of claim 1-10 composition, wherein the composition includes 102-1010Partial mature dendron
Cell.
13. any one of claim 1-10 composition, the ripe dendritic cells of which part are freezed after Partial mature
Preserve.
14. any one of claim 1-10 composition, it is used for the inducing antitumor immunity in the patient of this treatment of needs
Reaction, wherein, the composition is administered for the circulatory system or lymphatic system of patient.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US43126702P | 2002-12-06 | 2002-12-06 | |
| US60/431,267 | 2002-12-06 | ||
| CN2003801089756A CN1738619B (en) | 2002-12-06 | 2003-12-05 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2003801089756A Division CN1738619B (en) | 2002-12-06 | 2003-12-05 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102600461A CN102600461A (en) | 2012-07-25 |
| CN102600461B true CN102600461B (en) | 2017-07-14 |
Family
ID=32507696
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2003801089756A Expired - Fee Related CN1738619B (en) | 2002-12-06 | 2003-12-05 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
| CN201210027703.4A Expired - Fee Related CN102600461B (en) | 2002-12-06 | 2003-12-05 | Give outer body ripe dendritic cells treatment tumour |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2003801089756A Expired - Fee Related CN1738619B (en) | 2002-12-06 | 2003-12-05 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Country Status (19)
| Country | Link |
|---|---|
| US (2) | US20060057120A1 (en) |
| EP (2) | EP1567155B1 (en) |
| JP (2) | JP4859169B2 (en) |
| KR (1) | KR101144196B1 (en) |
| CN (2) | CN1738619B (en) |
| AT (1) | ATE486125T1 (en) |
| AU (2) | AU2003293411B2 (en) |
| BR (1) | BRPI0317064B8 (en) |
| CA (1) | CA2509058A1 (en) |
| DE (1) | DE60334725D1 (en) |
| DK (1) | DK1567155T3 (en) |
| ES (1) | ES2354944T3 (en) |
| HK (1) | HK1082180A1 (en) |
| IL (1) | IL169002A (en) |
| MX (1) | MXPA05006042A (en) |
| PL (1) | PL377209A1 (en) |
| PT (1) | PT1567155E (en) |
| RU (1) | RU2348418C2 (en) |
| WO (1) | WO2004053072A2 (en) |
Families Citing this family (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1567155B1 (en) * | 2002-12-06 | 2010-10-27 | NorthWest Biotherapeutics, Inc. | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
| EP1720567A2 (en) * | 2004-02-10 | 2006-11-15 | Innate Pharma | Composition and method for the treatment of carcinoma |
| US20060216269A1 (en) * | 2004-09-17 | 2006-09-28 | Kenichiro Hasumi | Dendritic cell tumor injection (DCTI) therapy |
| US8076132B2 (en) * | 2004-09-17 | 2011-12-13 | Hasumi International Research Foundation | Dendritic cell tumor injection (DCTI) therapy |
| WO2006095330A2 (en) * | 2005-03-10 | 2006-09-14 | Yeda Research And Development Co. Ltd. | Methods and immunogenic cell preparations for treating antigen-associated diseases |
| WO2007001200A1 (en) * | 2005-06-23 | 2007-01-04 | Obschestvo S Ogranichennoi Otvetsvennostyu 'rusgen' | Mature dendritic cells loaded with tumor polylysates and antitumoral vaccine base thereon |
| AR060424A1 (en) * | 2007-04-11 | 2008-06-18 | Consejo Nac Invest Cient Tec | CELLULAR LINES, COMPOSITIONS FOR THE TREATMENT OF MELANOMS THAT UNDERSTAND PROCEDURES TO PREPARE COMPOSITIONS AND TREATMENT METHODS |
| WO2008133595A1 (en) * | 2007-04-26 | 2008-11-06 | Newbiomed Pika Pte Ltd | Compositions and methods for stimulating an immune response in-vivo and in-vitro |
| WO2008151389A1 (en) * | 2007-06-15 | 2008-12-18 | Macfarlane Burnet Institute For Medical Research And Public Health Limited | Chemically modified macromolecules |
| EP2072617A1 (en) * | 2007-12-12 | 2009-06-24 | Trimed Biotech GmbH | Method for producing dendritic cells |
| JP5734836B2 (en) * | 2008-03-27 | 2015-06-17 | アステリアス バイオセラピューティクス インコーポレイテッド | Differentiation of primate pluripotent stem cells into hematopoietic lineage cells |
| NZ597975A (en) * | 2009-07-09 | 2014-06-27 | Tigenix S A | Methods and compositions for use in cellular therapies |
| JP5855018B2 (en) * | 2010-02-10 | 2016-02-09 | イミュニカム アーベー | Improved composition for inhibiting tumor cell growth |
| JP2013522309A (en) * | 2010-03-15 | 2013-06-13 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | System and method for preparing and storing activated mature dendritic cells |
| RU2530523C2 (en) * | 2012-03-29 | 2014-10-10 | Олег Борисович Егоров | Method of antitumour immunotherapy |
| EP3033417B1 (en) * | 2012-06-27 | 2019-12-25 | Kenichiro Hasumi | Therapy and method for intratumorally introducing cytotoxic t lymphocyte and/or nkt cell with anti-tnf and/or anti-il-10 |
| GB201300049D0 (en) * | 2013-01-03 | 2013-02-20 | Transimmune Ag | Method for obtaining immuno-stimulatory dendritic cells |
| BR112016010224A2 (en) | 2013-11-05 | 2018-05-02 | Cognate Bioservices, Inc. | checkpoint inhibitor combinations and therapeutic products to treat cancer. |
| GB201413665D0 (en) * | 2014-07-03 | 2014-09-17 | Transimmune Ag And Yale University | Method for obtaining globally activated monocytes |
| EP3317403A4 (en) | 2015-06-30 | 2019-07-03 | NorthWest Biotherapeutics, Inc. | Optimally activated dendritic cells that induce an improved or increased anti-tumor immune response |
| EP3350318B1 (en) * | 2015-09-15 | 2022-04-13 | NorthWest Biotherapeutics, Inc. | Methods relating to activated dendritic cell compositions for subjects with advanced cancers |
| KR101749165B1 (en) * | 2016-06-07 | 2017-06-23 | 충남대학교산학협력단 | Maturation compositions of dendritic cell containing the Rv2299c protein or Rv2299c fused ESAT-6 protein |
| WO2018160666A1 (en) * | 2017-02-28 | 2018-09-07 | University Of Pittsburgh-Of The Commonwealth System Of Higher Education | Short-term activated dc1s and methods for their production and use |
| GB201711379D0 (en) * | 2017-07-14 | 2017-08-30 | Univ Cape Town | Maturation of dendritic cells |
| CN109957548B (en) * | 2017-12-26 | 2022-03-18 | 上海尚泰生物技术有限公司 | Genetically modified dendritic cell vaccine |
| GB2624871A (en) * | 2022-11-29 | 2024-06-05 | Alv B As | Process for preparing a population of dendritic cells and immunotherapy using the same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1738619B (en) * | 2002-12-06 | 2012-04-18 | 西北生物治疗药物公司 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Family Cites Families (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2133409C (en) | 1992-04-01 | 2011-05-24 | Ralph M. Steinman | Method for in vitro proliferation of dendritic cell precursors and their use to produce immunogens |
| US5788963A (en) * | 1995-07-31 | 1998-08-04 | Pacific Northwest Cancer Foundation | Isolation and/or preservation of dendritic cells for prostate cancer immunotherapy |
| US6797488B1 (en) * | 1997-12-08 | 2004-09-28 | Beth Israel Deaconess Medical Center | Methods of producing anti-angiogenic proteins |
| EP1113821A1 (en) * | 1998-09-15 | 2001-07-11 | University Of Pittsburgh Of The Commonwealth System Of Higher Education | In situ injection of antigen-presenting cells with genetically enhanced cytokine expression |
| EP1126023A4 (en) * | 1998-10-02 | 2005-02-16 | Mitsubishi Chem Corp | Method for inducing cellular immunity and cells with induced cellular immunity |
| US6649158B1 (en) * | 1998-10-15 | 2003-11-18 | Canji, Inc. | Methods and compositions to induce antitumor response |
| US6558951B1 (en) * | 1999-02-11 | 2003-05-06 | 3M Innovative Properties Company | Maturation of dendritic cells with immune response modifying compounds |
| EP1171109A1 (en) * | 1999-04-14 | 2002-01-16 | THE GOVERNMENT OF THE UNITED STATES OF AMERICA, as represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES | Compositions containing immunotoxins and agents that inhibit dendritic cell maturation for inducing immune tolerance to a graft |
| US20030108527A1 (en) * | 1999-12-28 | 2003-06-12 | Tsukasa Seya | Maturation-promoting agent for immature dendrtic cells |
| JP2002069001A (en) * | 2000-08-29 | 2002-03-08 | Asahi Kasei Corp | Cell vaccine consisting mainly of dendritic cell |
| US20020094545A1 (en) | 2000-11-30 | 2002-07-18 | Harris Paul E. | Growth of human dendritic cells for cancer immunotherapy in closed system using microcarrier beads |
| WO2003010292A2 (en) * | 2001-07-25 | 2003-02-06 | Northwest Biotherapeutics, Inc. | Methods and apparatus for enrichment and culture of monocytic dendritic cell precursors |
| KR101803702B1 (en) * | 2001-09-06 | 2017-12-01 | 노쓰웨스트 바이오써라퓨틱스, 인크. | Compositions and methods for priming monocytic dendritic cells and T cells for Th-1 response |
| EP1604016B1 (en) * | 2003-02-27 | 2009-01-14 | NorthWest Biotherapeutics, Inc. | Generation of dendritic cells from monocytic dendritic precursor cells with gm-csf in the absence of additional cytokines |
-
2003
- 2003-12-05 EP EP03790358A patent/EP1567155B1/en not_active Expired - Lifetime
- 2003-12-05 CA CA002509058A patent/CA2509058A1/en not_active Expired - Lifetime
- 2003-12-05 ES ES03790358T patent/ES2354944T3/en not_active Expired - Lifetime
- 2003-12-05 CN CN2003801089756A patent/CN1738619B/en not_active Expired - Fee Related
- 2003-12-05 PL PL377209A patent/PL377209A1/en not_active Application Discontinuation
- 2003-12-05 US US10/538,226 patent/US20060057120A1/en not_active Abandoned
- 2003-12-05 PT PT03790358T patent/PT1567155E/en unknown
- 2003-12-05 DE DE60334725T patent/DE60334725D1/en not_active Expired - Lifetime
- 2003-12-05 MX MXPA05006042A patent/MXPA05006042A/en active IP Right Grant
- 2003-12-05 BR BRPI0317064A patent/BRPI0317064B8/en not_active IP Right Cessation
- 2003-12-05 CN CN201210027703.4A patent/CN102600461B/en not_active Expired - Fee Related
- 2003-12-05 EP EP20100176476 patent/EP2260861A1/en not_active Ceased
- 2003-12-05 WO PCT/US2003/038672 patent/WO2004053072A2/en active Application Filing
- 2003-12-05 JP JP2004559310A patent/JP4859169B2/en not_active Expired - Lifetime
- 2003-12-05 KR KR1020057010303A patent/KR101144196B1/en active IP Right Grant
- 2003-12-05 DK DK03790358.0T patent/DK1567155T3/en active
- 2003-12-05 RU RU2005121256/14A patent/RU2348418C2/en active IP Right Revival
- 2003-12-05 AT AT03790358T patent/ATE486125T1/en active
- 2003-12-05 AU AU2003293411A patent/AU2003293411B2/en not_active Ceased
-
2005
- 2005-06-05 IL IL169002A patent/IL169002A/en active IP Right Grant
-
2006
- 2006-02-02 HK HK06101427.5A patent/HK1082180A1/en not_active IP Right Cessation
-
2011
- 2011-01-28 AU AU2011200352A patent/AU2011200352B2/en not_active Ceased
- 2011-09-08 JP JP2011196385A patent/JP5707284B2/en not_active Expired - Fee Related
-
2012
- 2012-06-04 US US13/488,388 patent/US20120251561A1/en not_active Abandoned
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1738619B (en) * | 2002-12-06 | 2012-04-18 | 西北生物治疗药物公司 | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102600461B (en) | Give outer body ripe dendritic cells treatment tumour | |
| JP2006510667A5 (en) | ||
| JP2024101059A (en) | Optimally activated dendritic cells that induce improved or increased anti-tumor immune response | |
| RU2749610C2 (en) | Methods related to activated dendritic cells compositions and to immunotherapeutic treatment of individuals with advanced cancer | |
| AU2018260971B2 (en) | Administration of dendritic cells partially matured in vitro for the treatment of tumors | |
| AU2013234394B2 (en) | Administration of dendritic cells partially matured in vitro for the treatment of tumors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170714 Termination date: 20211205 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |